IgG Fc engineering to modulate antibody effector functions

Abstract

Therapeutic monoclonal antibodies are among the most effective biotherapeutics to date. An important aspect of antibodies is their ability to bind antigen while at the same time recruit immune effector functions. The majority of approved recombinant monoclonal antibody therapies are of the human IgG1 subclass, which can engage both humoral and cellular components of the immune system. The wealth of information generated about antibodies has afforded investigators the ability to molecularly engineer antibodies to modulate effector functions. Here, we review various antibody engineering efforts intended to improve efficacy and safety relative to the human IgG isotype. Further, we will discuss proposed mechanisms by which engineering approaches led to modified interactions with immune components and provide examples of clinical studies using next generation antibodies.

Key words： antibody-dependent cell-mediated cytotoxicity; antibody-dependent cellular phagocytosis; complement-dependent cytotoxicity; Fc engineering; Fc gamma receptor; monoclonal antibody; neonatal Fc receptor

References

1 Alegre ML, Collins AM, Pulito VL, Brosius RA, Olson WC, Zivin RA, Knowles R, Thistlethwaite JR, Jolliffe LK, Bluestone JA (1992) Effect of a single amino acid mutation on the activating and immunosuppressive properties of a “humanized” OKT3 monoclonal antibody. J Immunol 148:3461–3468

2 An Z, Forrest G, Moore R, Cukan M, Haytko P, Huang L, Vitelli S, Zhao JZ, Lu P, Hua J(2009) IgG2m4, an engineered antibody isotype with reduced Fc function. MAbs 1:572–579

3 Bang YJ, Giaccone G, Im SA, Oh DY, Bauer TM, Nordstrom JL, Li H, Chichili GR, Moore PA, Hong S (2017) First-in-human phase 1 study of margetuximab (MGAH22), an Fc-modified chimeric monoclonal antibody, in patients with HER2-positive advanced solid tumors. Ann Oncol 28:855–861

4 Beck A, Reichert JM (2012) Marketing approval of mogamulizumab: a triumph for glyco-engineering. MAbs 4:419–425

5 Bibeau F, Lopez-Crapez E, Di Fiore F, Thezenas S, Ychou M, Blanchard F, Lamy A, Penault-Llorca F, Frebourg T, Michel P(2009) Impact of Fc{gamma}RIIa-Fc{gamma}RIIIa polymorphisms and KRAS mutations on the clinical outcome of patients with metastatic colorectal cancer treated with cetuximab plus irinotecan. J Clin Oncol 27:1122–1129

6 Blank U, Launay P, Benhamou M, Monteiro RC (2009) Inhibitory ITAMs as novel regulators of immunity. Immunol Rev 232:59–71

7 Bolt S, Routledge E, Lloyd I, Chatenoud L, Pope H, Gorman SD, Clark M, Waldmann H (1993) The generation of a humanized, non-mitogenic CD3 monoclonal antibody which retains in vitro immunosuppressive properties. Eur J Immunol 23:403–411

8 Brambell FW (1966) The transmission of immunity from mother to young and the catabolism of immunoglobulins. Lancet 2:1087–1093

9 Brambell FW, Hemmings WA, Morris IG (1964) A theoretical model of gamma-globulin catabolism. Nature 203:1352–1354

10 Brezski RJ, Georgiou G (2016) Immunoglobulin isotype knowledge and application to Fc engineering. Curr Opin Immunol 40:62–69

11 Brezski RJ, Kinder M, Grugan KD, Soring KL, Carton J, Greenplate AR, Petley T, Capaldi D, Brosnan K, Emmell E (2014) A monoclonal antibody against hinge-cleaved IgG restores effector function to proteolytically-inactivated IgGs in vitro and in vivo. MAbs 6:1265–1273

12 Burmeister WP, Gastinel LN, Simister NE, Blum ML, Bjorkman PJ (1994a) Crystal structure at 2.2 A resolution of the MHC-related neonatal Fc receptor. Nature 372:336–343

13 Burmeister WP, Huber AH, Bjorkman PJ (1994b) Crystal structure of the complex of rat neonatal Fc receptor with Fc. Nature 372:379–383

14 Burton DR, Jefferis R, Partridge LJ, Woof JM (1988) Molecular recognition of antibody (IgG) by cellular Fc receptor (FcRI). Mol Immunol 25:1175–1181

15 Canfield SM, Morrison SL (1991) The binding affinity of human IgG for its high affinity Fc receptor is determined by multiple amino acids in the CH2 domain and is modulated by the hinge region. J Exp Med 173:1483–1491

16 Carter PJ (2006) Potent antibody therapeutics by design. Nat Rev Immunol 6:343–357

17 Cartron G, Dacheux L, Salles G, Solal-Celigny P, Bardos P, Colombat P, Watier H (2002) Therapeutic activity of humanized anti-CD20 monoclonal antibody and polymorphism in IgG Fc receptor FcgammaRIIIa gene. Blood 99:754–758

18 Chappel MS, Isenman DE, Everett M, Xu YY, Dorrington KJ, Klein MH (1991) Identification of the Fc gamma receptor class I binding site in human IgG through the use of recombinant IgG1/IgG2 hybrid and point-mutated antibodies. Proc Natl Acad Sci USA 88:9036–9040

19 Chatenoud L, Bluestone JA (2007) CD3-specific antibodies: a portal to the treatment of autoimmunity. Nat Rev Immunol 7:622–632

20 Chu SY, Vostiar I, Karki S, Moore GL, Lazar GA, Pong E, Joyce PF, Szymkowski DE, Desjarlais JR (2008) Inhibition of B cell receptor-mediated activation of primary human B cells by coengagement of CD19 and FcgammaRIIb with Fc-engineered antibodies. Mol Immunol 45:3926–3933

21 Dall’Acqua WF, Woods RM, Ward ES, Palaszynski SR, Patel NK, Brewah YA, Wu H, Kiener PA, Langermann S (2002) Increasing the affinity of a human IgG1 for the neonatal Fc receptor: biological consequences. J Immunol 169:5171–5180

22 Dall’Acqua WF, Kiener PA, Wu H (2006) Properties of human IgG1s engineered for enhanced binding to the neonatal Fc receptor (FcRn). J Biol Chem 281:23514–23524

23 Diebolder CA, Beurskens FJ, de Jong RN, Koning RI, Strumane K, Lindorfer MA, Voorhorst M, Ugurlar D, Rosati S, Heck AJ (2014) Complement is activated by IgG hexamers assembled at the cell surface. Science 343:1260–1263

24 Duncan AR, Woof JM, Partridge LJ, Burton DR, Winter G (1988) Localization of the binding site for the human high-affinity Fc receptor on IgG. Nature 332:563–564

25 Ecker DM, Jones SD, Levine HL (2015) The therapeutic monoclonal antibody market. MAbs 7:9–14

26 Edelman GM, Cunningham BA, Gall WE, Gottlieb PD, Rutishauser U, Waxdal MJ (1969) The covalent structure of an entire gammaG immunoglobulin molecule. Proc Natl Acad Sci USA 63:78–85

27 Fahey JL, Robinson AG (1963) Factors controlling serum gammaglobulin concentration. J Exp Med 118:845–868

28 Ferrara C, Grau S, Jager C, Sondermann P, Brunker P, Waldhauer I, Hennig M, Ruf A, Rufer AC, Stihle M (2011) Unique carbohydrate-carbohydrate interactions are required for high affinity binding between FcgammaRIII and antibodies lacking core fucose. Proc Natl Acad Sci USA 108:12669–12674

29 Gavin PG, Song N, Kim SR, Lipchik C, Johnson NL, Bandos H, Finnigan M, Rastogi P, Fehrenbacher L, Mamounas EP (2017) Association of polymorphisms in FCGR2A and FCGR3A with degree of trastuzumab benefit in the adjuvant treatment of ERBB2/HER2-positive breast cancer: analysis of the NSABP B-31 trial. JAMA Oncol 3:335–341

30 Ghetie V, Hubbard JG, Kim JK, Tsen MF, Lee Y, Ward ES (1996) Abnormally short serum half-lives of IgG in beta 2-microglobulindeficient mice. Eur J Immunol 26:690–696

31 Hulett MD, Witort E, Brinkworth RI, McKenzie IF, Hogarth PM (1994) Identification of theIgGbinding site of the human lowaffinity receptor for IgGFc gamma RII. Enhancement and ablation of binding by sitedirected mutagenesis. J Biol Chem 269:15287–15293

32 Idusogie EE, Presta LG, Gazzano-Santoro H, Totpal K, Wong PY, Ultsch M, Meng YG, Mulkerrin MG (2000) Mapping of the C1q binding site on rituxan, a chimeric antibody with a human IgG1 Fc. J Immunol 164:4178–4184

33 Idusogie EE, Wong PY, Presta LG, Gazzano-Santoro H, Totpal K, Ultsch M, Mulkerrin MG (2001) Engineered antibodies with increased activity to recruit complement. J Immunol 166:2571–2575

34 Israel EJ, Wilsker DF, Hayes KC, Schoenfeld D, Simister NE (1996) Increased clearance of IgG in mice that lack beta 2-microglobulin: possible protective role of FcRn. Immunology 89:573–578

35 Jefferis R, Lund J, Pound J (1990) Molecular definition of interaction sites on human IgG for Fc receptors (huFc gamma R). Mol Immunol 27:1237–1240

36 Junghans RP, Anderson CL (1996) The protection receptor for IgG catabolism is the beta2-microglobulin-containing neonatal intestinal transport receptor. Proc Natl Acad Sci USA 93:5512–5516

37 Kim JK, Firan M, Radu CG, Kim CH, Ghetie V, Ward ES (1999) Mapping the site on human IgG for binding of the MHC class I-related receptor, FcRn. Eur J Immunol 29:2819–2825

38 Kiyoshi M, Caaveiro JM, Kawai T, Tashiro S, Ide T, Asaoka Y, Hatayama K, Tsumoto K (2015) Structural basis for binding of human IgG1 to its high-affinity human receptor FcgammaRI. Nat Commun 6:6866

39 Lazar GA, Dang W, Karki S, Vafa O, Peng JS, Hyun L, Chan C, Chung HS, Eivazi A, Yoder SC (2006) Engineered antibody Fc variants with enhanced effector function. Proc Natl Acad Sci USA 103:4005–4010

40 Leabman MK, Meng YG, Kelley RF, DeForge LE, Cowan KJ, Iyer S (2013) Effects of altered FcgammaR binding on antibody pharmacokinetics in cynomolgus monkeys. MAbs 5:896–903

41 Lefran MPL (2001) The immunoglobulin factsbook. Elsevier, Amsterdam

42 Liu H, May K (2012) Disulfide bond structures of IgG molecules: structural variations, chemical modifications and possible impacts to stability and biological function. MAbs 4:17–23

43 Lo M, Kim HS, Tong RK, Bainbridge TW, Vernes JM, Zhang Y, Lin YL, Chung S, Dennis MS, Zuchero YJ (2017) Effectorattenuating substitutions that maintain antibody stability and reduce toxicity in mice. J Biol Chem 292:3900–3908

44 Loyau J, Malinge P, Daubeuf B, Shang L, Elson G, Kosco-Vilbois M, Fischer N, Rousseau F (2014) Maximizing the potency of an anti-TLR4 monoclonal antibody by exploiting proximity to Fcgamma receptors. MAbs 6:1621–1630

45 Lu J, Chu J, Zou Z, Hamacher NB, Rixon MW, Sun PD (2015) Structure of FcgammaRI in complex with Fc reveals the importance of glycan recognition for high-affinity IgG binding. Proc Natl Acad Sci USA 112:833–838

46 Lund J, Winter G, Jones PT, Pound JD, Tanaka T, Walker MR, Artymiuk PJ, Arata Y, Burton DR, Jefferis R (1991) Human Fc gamma RI and Fc gamma RII interact with distinct but overlapping sites on human IgG. J Immunol 147:2657–2662

47 Martin WL, West AP Jr, Gan L, Bjorkman PJ (2001) Crystal structure at 2.8 A of an FcRn/heterodimeric Fc complex: mechanism of pHdependent binding. Mol Cell 7:867–877

48 Medesan C, Matesoi D, Radu C, Ghetie V, Ward ES (1997) Delineation of the amino acid residues involved in transcytosis and catabolism of mouse IgG1. J Immunol 158:2211–2217

49 Meyer S, Leusen JH, Boross P (2014) Regulation of complement and modulation of its activity in monoclonal antibody therapy of cancer. MAbs 6:1133–1144

50 Mimoto F, Igawa T, Kuramochi T, Katada H, Kadono S, Kamikawa T, Shida-Kawazoe M, Hattori K (2013a) Novel asymmetrically engineered antibody Fc variant with superior FcgammaR binding affinity and specificity compared with afucosylated Fc variant. MAbs 5:229–236

51 Mimoto F,Katada H, Kadono S, Igawa T, Kuramochi T, Muraoka M, Wada Y, Haraya K, Miyazaki T, Hattori K (2013b) Engineered antibody Fc variant with selectively enhanced FcgammaRIIb binding over both FcgammaRIIa(R131) and FcgammaRIIa (H131). Protein Eng Des Sel 26:589–598

52 Monnet E, Lapeyre G, Poelgeest EV, Jacqmin P, Graaf K, Reijers J, Moerland M, Burggraaf J, Min C (2017) Evidence of NI-0101 pharmacological activity, an anti-TLR4 antibody, in a randomized phase I dose escalation study in healthy volunteers receiving LPS. Clin Pharmacol Ther 101:200–208

53 Moore GL, Chen H, Karki S, Lazar GA (2010) Engineered Fc variant antibodies with enhanced ability to recruit complement and mediate effector functions. MAbs 2:181–189

54 Morell A, Terry WD, Waldmann TA (1970) Metabolic properties of IgG subclasses in man. J Clin Invest 49:673–680

55 Natsume A, In M, Takamura H, Nakagawa T, Shimizu Y, Kitajima K, Wakitani M, Ohta S, Satoh M, Shitara K (2008) Engineered antibodies of IgG1/IgG3 mixed isotype with enhanced cytotoxic activities. Cancer Res 68:3863–3872

56 Nesspor TC, Raju TS, Chin CN, Vafa O, Brezski RJ (2012) Avidity confers FcgammaR binding and immune effector function to aglycosylated immunoglobulin G1. J Mol Recognit 25:147–154

57 Nimmerjahn F, Ravetch JV (2008) Fcgamma receptors as regulators of immune responses. Nat Rev Immunol 8:34–47

58 Oganesyan V, Damschroder MM, Leach W, Wu H, Dall’Acqua WF (2008) Structural characterization of a mutated, ADCC-enhanced human Fc fragment. Mol Immunol 45:1872–1882

59 Oganesyan V, Damschroder MM, Cook KE, Li Q,Gao C, Wu H, Dall’Acqua WF(2014) Structural insights into neonatal Fc receptorbased recycling mechanisms. J Biol Chem 289:7812–7824

60 Oganesyan V, Mazor Y, Yang C, Cook KE, Woods RM, Ferguson A, Bowen MA, Martin T, Zhu J, Wu H (2015) Structural insights into the interaction of human IgG1 with FcgammaRI: no direct role of glycans in binding. Acta Crystallogr D 71:2354–2361

61 Partridge LJ, Woof JM, Jefferis R, Burton DR (1986) The use of anti-IgG monoclonal antibodies in mapping the monocyte receptor site on IgG. Mol Immunol 23:1365–1372

62 Presta LG (2008) Molecular engineering and design of therapeutic antibodies. Curr Opin Immunol 20:460–470

63 Radaev S, Motyka S, Fridman WH, Sautes-Fridman C, Sun PD (2001) The structure of a human type III Fcgamma receptor in complex with Fc. J Biol Chem 276:16469–16477

64 Ramsland PA, Farrugia W, Bradford TM, Sardjono CT, Esparon S,Trist HM, Powell MS, Tan PS, Cendron AC, Wines BD (2011) Structural basis for Fc gammaRIIa recognition of human IgG and formation of inflammatory signaling complexes. J Immunol 187:3208–3217

65 Rath T, Kuo TT, Baker K, Qiao SW, Kobayashi K, Yoshida M, Roopenian D, Fiebiger E, Lencer WI, Blumberg RS (2013) The immunologic functions of the neonatal Fc receptor for IgG. J Clin Immunol 33(Suppl 1):S9–17

66 Reichert JM (2017) Antibodies to watch in 2017. MAbs 9:167–181

67 Richards JO, Karki S, Lazar GA, Chen H, Dang W, Desjarlais JR (2008) Optimization of antibody binding to FcgammaRIIa enhances macrophage phagocytosis of tumor cells. Mol Cancer Ther 7:2517–2527

68 Robbie GJ, Criste R, Dall’acqua WF, Jensen K, Patel NK, Losonsky GA, Griffin MP (2013) A novel investigational Fc-modified humanized monoclonal antibody, motavizumab-YTE, has an extended half-life in healthy adults. Antimicrob Agents Chemother 57:6147–6153

69 Roopenian DC, Akilesh S (2007) FcRn: the neonatal Fc receptor comes of age. Nat Rev Immunol 7:715–725

70 Rother RP, Rollins SA, Mojcik CF, Brodsky RA, Bell L (2007) Discovery and development of the complement inhibitor eculizumab for the treatment of paroxysmal nocturnal hemoglobinuria. Nat Biotechnol 25:1256–1264

71 Shang L,Daubeuf B, Triantafilou M,Olden R, Depis F, Raby AC, Herren S, Dos Santos A, Malinge P, Dunn-Siegrist I (2014) Selective antibody intervention of Toll-like receptor 4 activation through Fc gamma receptor tethering. J Biol Chem 289:15309–15318

72 Shields RL, Namenuk AK, Hong K, Meng YG, Rae J, Briggs J, Xie D, Lai J, Stadlen A, Li B (2001) High resolution mapping of the binding site on human IgG1 for Fc gamma RI, Fc gamma RII, Fc gamma RIII, and FcRn and design of IgG1 variants with improved binding to the Fc gamma R. J Biol Chem 276:6591–6604

73 Shields RL, Lai J, Keck R, O’Connell LY, Hong K, Meng YG, Weikert SH, Presta LG (2002) Lack of fucose on human IgG1 N-linked oligosaccharide improves binding to human Fcgamma RIII and antibody-dependent cellular toxicity. J Biol Chem 277:26733–26740

74 Shinkawa T, Nakamura K, Yamane N, Shoji-Hosaka E, Kanda Y, Sakurada M, Uchida K, Anazawa H, Satoh M, Yamasaki M (2003) The absence of fucose but not the presence of galactose or bisecting N-acetylglucosamine of human IgG1 complex-type oligosaccharides shows the critical role of enhancing antibody-dependent cellular cytotoxicity. J Biol Chem 278:3466–3473

75 Sondermann P, Huber R, Oosthuizen V, Jacob U (2000) The 3.2-A crystal structure of the human IgG1 Fc fragment-Fc gammaRIII complex. Nature 406:267–273

76 Stavenhagen JB, Gorlatov S, Tuaillon N, Rankin CT, Li H, Burke S, Huang L, Vijh S, Johnson S, Bonvini E (2007) Fc optimization of therapeutic antibodies enhances their ability to kill tumor cells in vitro and controls tumor expansion in vivo via low-affinity activating Fcgamma receptors. Cancer Res 67:8882–8890

77 Strohl WR (2011) Isotype selection and Fc engineering: design and construction of fit-for-purpose therapeutic antibodies. Antibody drug discovery Imperial College Press, London

78 Strohl WR (2017) Current progress in innovative engineered antibodies. Protein Cell.

79 Szili D, Cserhalmi M, Banko Z, Nagy G, Szymkowski DE, Sarmay G (2014) Suppression of innate and adaptive B cell activation pathways by antibody coengagement of FcgammaRIIb and CD19. MAbs 6:991–999

80 Tamm A, Kister A, Nolte KU, Gessner JE, Schmidt RE (1996) The IgG binding site of human FcgammaRIIIB receptor involves CC’ and FG loops of the membrane-proximal domain. J Biol Chem 271:3659–3666

81 Tao MH, Morrison SL (1989) Studies of aglycosylated chimeric mouse-human IgG. Role of carbohydrate in the structure and effector functions mediated by the human IgG constant region. J Immunol 143:2595–2601

82 Tao MH, Smith RI, Morrison SL (1993) Structural features of human immunoglobulin G that determine isotype-specific differences in complement activation. J Exp Med 178:661–667

83 Teeling JL, Mackus WJ, Wiegman LJ, van den Brakel JH, Beers SA, French RR, van Meerten T, Ebeling S, Vink T, Slootstra JW (2006) The biological activity of human CD20 monoclonal antibodies is linked to unique epitopes on CD20. J Immunol 177:362–371

84 Thommesen JE, Michaelsen TE, Loset GA, Sandlie I, Brekke OH (2000) Lysine 322 in the human IgG3 C(H)2 domain is crucial for antibody dependent complement activation. Mol Immunol 37:995–1004

85 Umana P, Jean-Mairet J, Moudry R, Amstutz H, Bailey JE (1999) Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity. Nat Biotechnol 17:176–180

86 Uppal H, Doudement E, Mahapatra K, Darbonne WC, Bumbaca D, Shen BQ, Du X, Saad O, Bowles K, Olsen S (2015) Potential mechanisms for thrombocytopenia development with trastuzumab emtansine (T-DM1). Clin Cancer Res 21:123–133

87 Vafa O,Gilliland GL, Brezski RJ, Strake B, Wilkinson T, Lacy ER, Scallon B, Teplyakov A, Malia TJ, Strohl WR (2014) An engineered Fc variant of an IgG eliminates all immune effector functions via structural perturbations. Methods 65:114–126

88 Walker MR, Lund J, Thompson KM, Jefferis R (1989) Aglycosylation of human IgG1 and IgG3 monoclonal antibodies can eliminate recognition by human cells expressing Fc gamma RI and/or Fc gamma RII receptors. Biochem J 259:347–353

89 Wines BD, Powell MS, Parren PW, Barnes N, Hogarth PM (2000) The IgG Fc contains distinct Fc receptor (FcR) binding sites: the leukocyte receptors Fc gamma RI and Fc gamma RIIa bind to a region in the Fc distinct from that recognized by neonatal FcR and protein A. J Immunol 164:5313–5318

90 Woof JM, Partridge LJ, Jefferis R, Burton DR (1986) Localisation of the monocyte-binding region on human immunoglobulin G. Mol Immunol 23:319–330

91 Xu D, Alegre ML, Varga SS, Rothermel AL, Collins AM, Pulito VL, Hanna LS, Dolan KP, Parren PW, Bluestone JA (2000) In vitro characterization of five humanized OKT3 effector function variant antibodies. Cell Immunol 200:16–26

92 Zalevsky J, Chamberlain AK, Horton HM, Karki S, Leung IW, Sproule TJ, Lazar GA, Roopenian DC, Desjarlais JR (2010) Enhanced antibody half-life improves in vivo activity. Nat Biotechnol 28:157–159